Journal: Journal of Neuroinflammation

Article Title: Apolipoprotein E isoform dependently affects Tat-mediated HIV-1 LTR transactivation

doi: 10.1186/s12974-018-1129-1

Figure Lengend Snippet: Involvement of LRP1 in Tat-mediated HIV-1 LTR transactivation. a The LRP1 antagonist, receptor-associated protein (RAP), concentration dependently reduced Tat-mediated HIV-1 LTR transactivation ( n = 3; * p < 0.05). b Protein expression levels of LRP1 were significantly ( n = 3, p < 0.05) reduced in U87MG cells transfected with LRP1 siRNA (100 nM) compared to cells transfected with control (scrambled) siRNA. c siRNA knockdown of LRP1 reduced significantly ( n = 3; * p < 0.05) Tat-mediated HIV-1 LTR transactivation

Article Snippet: Cells plated on glass-bottom 35-mm 2 tissue culture dishes were co-incubated with ApoE2, ApoE3, or ApoE4 (5,10 μg/ml) and FITC-labeled HIV-1 Tat (5 μg/ml; purchased from Rockland) for 90 min at 37 °C in the presence of chloroquine (100 μM).

Techniques: Concentration Assay, Expressing, Transfection

Journal: Journal of Neuroinflammation

Article Title: Apolipoprotein E isoform dependently affects Tat-mediated HIV-1 LTR transactivation

doi: 10.1186/s12974-018-1129-1

Figure Lengend Snippet: ApoE-HDL isoform dependently affected Tat-mediated HIV-1 LTR transactivation. U87MG cells stably transfected with a luciferase gene under the control of HIV-1 Tat responsive LTR promoter were incubated for 48 h with ApoE2-HDL ( a ), ApoE3-HDL ( b ), or ApoE4-HDL ( c ) in the presence of HIV-1 Tat protein (2 μg/ml) and 100 μM chloroquine (CQ). ApoE2-HDL, ApoE3-HDL, and ApoE4-HDL significantly and concentration dependently restricted Tat-mediated HIV-1 LTR transactivation ( n = 3; * p < 0.05, ** p < 0.01, *** p < 0.001) ( d ). In comparison to ApoE2-HDL and ApoE3-HDL, ApoE4-HDL was less potent and effective at restricting Tat-mediated HIV-1 transactivation

Article Snippet: Cells plated on glass-bottom 35-mm 2 tissue culture dishes were co-incubated with ApoE2, ApoE3, or ApoE4 (5,10 μg/ml) and FITC-labeled HIV-1 Tat (5 μg/ml; purchased from Rockland) for 90 min at 37 °C in the presence of chloroquine (100 μM).

Techniques: Stable Transfection, Transfection, Luciferase, Incubation, Concentration Assay

Journal: Journal of Neuroinflammation

Article Title: Apolipoprotein E isoform dependently affects Tat-mediated HIV-1 LTR transactivation

doi: 10.1186/s12974-018-1129-1

Figure Lengend Snippet: HDL affects the ability of ApoE to restrict Tat-mediated HIV-1 LTR transactivation. a ApoE2 significantly and concentration dependently restricted Tat-mediated HIV-1 LTR transactivation in the absence of HDL ( n = 3; * p < 0.05; ** p < 0.01). b ApoE3 significantly and concentration dependently restricted Tat-mediated HIV-1 LTR transactivation in the absence of HDL ( n = 3; ** p < 0.01). c ApoE4 concentration dependently enhanced Tat-mediated HIV-1 LTR transactivation in the absence of HDL ( n = 3, ** p < 0.01). d In the presence of HDL, ApoE2 was more potent and effective at restricting Tat-mediated HIV-1 LTR transactivation. e In the presence of HDL, ApoE3 was more potent and effective at restricting Tat-mediated HIV-1 LTR transactivation. f In presence of HDL, ApoE4 concentration dependently restricted Tat-mediated HIV-1 LTR transactivation; however, in the absence of HDL, ApoE4 concentration dependently enhanced Tat-mediated HIV-1 LTR transactivation

Article Snippet: Cells plated on glass-bottom 35-mm 2 tissue culture dishes were co-incubated with ApoE2, ApoE3, or ApoE4 (5,10 μg/ml) and FITC-labeled HIV-1 Tat (5 μg/ml; purchased from Rockland) for 90 min at 37 °C in the presence of chloroquine (100 μM).

Techniques: Concentration Assay

Journal: Journal of Neuroinflammation

Article Title: Apolipoprotein E isoform dependently affects Tat-mediated HIV-1 LTR transactivation

doi: 10.1186/s12974-018-1129-1

Figure Lengend Snippet: ApoE2 and ApoE3, but not ApoE4, inhibited HIV-1 Tat internalization. Representative images ( a ) and quantified internalized fluorescence intensities ( b ) show that ApoE2 and ApoE3 at 5 and 10 μg/ml inhibited HIV-1 Tat internalization as evidenced by lower levels of punctate staining of internalized FITC-labeled Tat ( n = 30; *** p < 0.001, **** p < 0.0001), whereas ApoE4 did not inhibit HIV-1 Tat internalization

Article Snippet: Cells plated on glass-bottom 35-mm 2 tissue culture dishes were co-incubated with ApoE2, ApoE3, or ApoE4 (5,10 μg/ml) and FITC-labeled HIV-1 Tat (5 μg/ml; purchased from Rockland) for 90 min at 37 °C in the presence of chloroquine (100 μM).

Techniques: Fluorescence, Staining, Labeling

Journal: Journal of Neuroinflammation

Article Title: Apolipoprotein E isoform dependently affects Tat-mediated HIV-1 LTR transactivation

doi: 10.1186/s12974-018-1129-1

Figure Lengend Snippet: ApoE mimetic peptide decreased Tat-mediated HIV-1 LTR transactivation. a U87MG cells were treated with an ApoE mimetic peptide in the presence of HIV-1 Tat and chloroquine (CQ) for 48 h. ApoE mimetic peptide significantly reduced Tat-mediated HIV-1 LTR transactivation ( n = 3; * p < 0.05). b Incubation with an ApoE4 structure corrector, that makes the structure and function of ApoE4 more like ApoE3, enhanced the ability of ApoE4 to restrict Tat-mediated LTR transactivation ( n = 3; * p < 0.05)

Article Snippet: Cells plated on glass-bottom 35-mm 2 tissue culture dishes were co-incubated with ApoE2, ApoE3, or ApoE4 (5,10 μg/ml) and FITC-labeled HIV-1 Tat (5 μg/ml; purchased from Rockland) for 90 min at 37 °C in the presence of chloroquine (100 μM).

Techniques: Incubation

Journal: Scientific Reports

Article Title: Potential Orphan Drug Therapy of Intravesical Liposomal Onabotulinumtoxin-A for Ketamine-Induced Cystitis by Mucosal Protection and Anti-inflammation in a Rat Model

doi: 10.1038/s41598-018-24239-9

Figure Lengend Snippet: Alterations of neuroreceptors protein expression in the mucosa layer or smooth muscle layer of the bladder for all groups (n = 8). Western blot analyses with specific antibodies to the TRPV1 receptor and P2X 3 receptor of the rat mucosal layer as well as M 2 -and M 3 - mAChRs and the purinergic P2X 1 receptor of the rat detrusor layer were performed in 3 groups. ( A ) TRPV1 receptor: The TRPV1 antibody produced a clear single band at 95 kDa. ( B ) Purinergic P2X 3 mature receptor: the predominant P2X 3 form (65 kDa). ( C ) M 2 –mAChR of bladder detrusor layer. The M 2 –mAChR antibody produced a clear single band between 50kD and 75kD. ( D ) M 3 –mAChR of bladder detrusor layer. ( E ) Purinergic P2X 1 receptor. Experiments were repeated two times and representative blots are shown. Data of proteins expression (ratios of signal intensities of investigated receptors relative to GAPDH) were calculated with 8 samples in each group. These data of Mean ± SE were standardized and expressed in percentage in which the value of the control group is treated as 100%. Theses values were shown in the bar graph. An asterisk indicates a significant difference between controls and other groups (One-way ANOVA with Dunnett’s test, p < 0.05). The grouping of blots was cropped from the same gel for each protein. The full-length gels and blots are included in the Supplementary Figure .

Article Snippet: Antibodies raised against SNAP25 (1: 1000 dilution; Cell signal), E-cadherin (1:1000 dilution; Cell Signal), nerve growth factor (1:200 dilution; Cell Signal), IL-1β (1:500 dilution; Abcam) IL-6 (1:1000 dilution; Abcam), TNF-α (1:250 dilution, Santa Cruz), NF-κB (1: 2500 dilution; Cell SiZgnal), COX-2 (1:200 dilution; Abcam), TRPV1 receptor (1:1000 dilution; Alomone), purinergic receptor P2X 1 (1:10000 dilution; Alomone), purinergic receptor P2X 3 (1: 1000 dilution; Alomone), M 2 -mAChR (1:1000 dilution; Alomone), M 3 -mAChR (1:1000 dilution; Alomone), and GAPDH (1: 10,000 dilution; Millipore) were used.

Techniques: Expressing, Western Blot, Produced